LOCUS       sequence-436659-        5378 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector BS2 in nucleolus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5378)
  AUTHORS   Pani S, Qiu T, Kentala K, Azizi SA, Dickinson BC
  TITLE     Bioorthogonal masked acylating agents for proximity-dependent RNA 
            labelling.
  JOURNAL   Nat Chem. 2024 Apr 9. doi: 10.1038/s41557-024-01493-1.
  PUBMED    38594368
REFERENCE   2  (bases 1 to 5378)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Chem. 
            2024 Apr 9. doi: 10.1038/s41557-024-01493-1."
COMMENT     Sequence Label: V5-BS2-NIK3x (Nucleolus)
FEATURES             Location/Qualifiers
     source          1..5378
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(439..480)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     regulatory      480..489
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(531..550)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          533..665
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(801..1004)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(830..850)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1005..1384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    1575..1858
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     rep_origin      1974..2562
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2463..2482
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2734..3528)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFAKLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2845..2864)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3455..3474
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     polyA_signal    3927..4048
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3964..3983)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4018..4037
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4085..4103
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 agacaagtgc aaaaagccaa acggccccag ccgatagttc aatgtaacga caatgacttc
       61 tccctgtgcc gcaagttttg atccgtcata caatggctca ctgcccgctc ctagataaaa
      121 agcgcctccg tgaatccaca ccatgacagg aagattttta cttggggtgt caggcgcaaa
      181 tacattgaca tacaagcaat cctcggactg gcggggcagc tcagtatacg aaagtgacag
      241 caaatcagac ggctgcgggc aaatagagcc gtacgctgtg gcatcaagca catcttccca
      301 cacttcaggc ggctcaggtg ctttaaaacg ccattgtccg acaggcggct tggcataggg
      361 gatgcctttc cacttatgta cgccgttttc cgttgtgcct tttactttgc cgtattgagt
      421 cgttactatt tgatgagtgg tgctgtccag gcccagcagg gggttgggga tgggcttgcc
      481 catggtggca tactctagcc ttaagagctg taattgaact gggagtggac acctgtggag
      541 agaaaggcaa agtggatgtc agtaagacca ataggtgcct atcagaaacg caagagtctt
      601 ctctgtctcg acaagcccag tttctattgg tctccttaaa cctgtcttgt aaccttgata
      661 cttacctgcc cagtgcctca cgaccaactt ctgcagctta agttcgagac tgttgtgtca
      721 gaagcactga ctgcgttagc aatttaactg tgataaacta ccgcaataaa gcttctagtg
      781 atctgacggt tcactaaacc agctctgctt atatagacct cccaccgtac acgcctaccg
      841 cccatttgcg tcaacggggc ggggttatta cgacattttg gaaagtcccg ttgattttgg
      901 tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca
      961 aaccgctatc cacgcccatt ggtgtactgc caaaaccgca tcaccatggt aatagcgatg
     1021 actaatacgt agatgtactg ccaagtagga aagtcccgta aggtcatgta ctgggcataa
     1081 tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcggacttgg catatgatac
     1141 acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg
     1201 aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg
     1261 tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata
     1321 tatgggctat gaactaatga ccccgtaatt gattactatt aataactagt caataatcaa
     1381 tgccaacatg gcggtcatat tggacatgag ccaatataaa tgtacatatt atgatataga
     1441 tacaacgtat gcaatggcca atagccaata ttgatttatg ctatataacc aatgaataat
     1501 atggctaatg gccaatattg aagatctcga tcctctacgc cggacgcatc gtggccggag
     1561 tactgcggtt tatctcgatg gctacgaggg cagacagtaa gtggatttac cataatccct
     1621 taattgtacg caccgctaaa acgcgttcag cgcgatcacg gcagcagaca ggtaaaaatg
     1681 gcaacaaacc acccgaaaaa ctgccgcaat cgcgcctgat aaattttaac cgtatgaata
     1741 cctatgcaac cagagggtac aggccacatt acccccactt aatccactga agctgccatt
     1801 tttcatggtt tcaccatccc agcgaagggc catccagcgt gcgttcctgt atttccggac
     1861 tataaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cacttaacgt
     1921 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgaaat
     1981 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2041 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2101 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     2161 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2221 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2281 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2341 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     2401 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     2461 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2521 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2581 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     2641 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgg ttgggcgtcg
     2701 cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa gaaggcgata
     2761 gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga agcggtcagc
     2821 ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt cctgatagcg
     2881 gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat tttccaccat
     2941 gatattcggc aagcaggcat cgccatgagt cacgacgaga tcctcaccat ccggcatacg
     3001 cgccttgagc ttggcgaaca gttcggctgg cgcgagcccc tgatgctctt cgtccagatc
     3061 atcctgatcg acaagaccgg cttccatccg agtgcgtgct cgctcgatgc gatgtttcgc
     3121 ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca ttgcatcagc
     3181 catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct gccccggcac
     3241 ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca cagctgcgca
     3301 aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgca gttcattcag
     3361 ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa
     3421 cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga atagcctctc
     3481 cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaagcatgc taaacgaccg
     3541 tcatcctgtc tcttgatcag atcttgatcc cctgcgccat cagatccttg gcggcaagaa
     3601 agccatccag tttactttgc agggcttccc aaccttacca gagggcgccc cagctggcaa
     3661 ttcttttgaa gctcacgctg ccgcaagcac tcagggcgca agggctgcta aaggaagcgg
     3721 aacacgtaga aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg
     3781 gctatctgga caagggaaaa cgcaagcgca aagaggatcc attagaattc gattttacca
     3841 catttgtaga ggttttactt gcttaaaaaa acctcccaca tctccccctg aacctgaaac
     3901 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3961 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4021 gtttgtccaa actcatcaat gtatcttatc atgtctgctc gaagcggccg ccccaagggg
     4081 ttatgctagt tattgctcag cggtggcagc agccaactca gcttcctttc gggctttgtt
     4141 agcagccgga tcagcttgca tgcctgcagg tcgactctag aggatccccg ggtaccgagc
     4201 ccgaattcgt ttaaacatca caggatgcgc agcttcttct tgcgtttctt gcgcatcctg
     4261 aggatgcgga gcttcttctt gcgcttcttt ctcatgcgca ggattcgaag cttctttttt
     4321 cgcttctttc tcattcttga attcgaggat ccttctcctt ttgaagggaa tagcttctgc
     4381 cttttttcag attcgggatc gttttcgatc gtaatctctg agtctaaaat cagcgtctct
     4441 ctcgtttctt catgatacgc aggccaattc acagcttcgg tgcttgggtt tcctgttttg
     4501 gcgaacgtga tccacgctga ttgtatcgtg tgagaaagct gtttcacctc atccgtaatc
     4561 tccgcttttg ccattcgttc caatccgtcc agatttccaa agacaaaagg aagctctaat
     4621 gcgtgaaacg ctttattgta cggcggcttc ttcgggtgcc aatcgaacct gtacatccag
     4681 acaggggcgt aatgagactg tgcggatgca taggcgacgg cagggcgcca aaataataaa
     4741 tcagtcatca tgtgaatttg gctttccaga gaacgcggat acaaatcggc aactttctct
     4801 gccagcggct tccctagtaa atactcgagc gctgcatcaa gcgtttcctg agaatgaacg
     4861 tctgaatccg gggtgaaaaa taaatatcct tcatcacggg ttgttccaat taatagcgga
     4921 ataccggaag cagccccttc tgcgatcgct ttttctggtt cttcaggcag cgttttcgga
     4981 tcaagggcgg gctggaagaa cagctgaaag atattttctt tttctgcaat ccgaagctga
     5041 tcagccgctt ttagcaaatc ttccgcagaa accgtatgca atttatccag ttggccctcg
     5101 ttaatcccaa ggacctgtaa aaaggctgcc gaggtgctcg ccgcttgttc tttcgtcatc
     5161 gttcgagaag cgccgctttc catgattgct ttctggaaca ggccttttgc cgcaggcata
     5221 gcaagcagcg cggcaatgct catcccgccg gcggattctc caaatactgt tacgttatcg
     5281 ggatcaccgc caaacgctga aatattctct cgcacccatt tcagcgcggc ggcttggtct
     5341 aaaagcccaa ggttatcaga atacgcctca ttaaatga
//
